UMMS Affiliation

Department of Molecular, Cell and Cancer Biology; Department of Biochemistry and Molecular Pharmacology; UMass Metabolic Network

Publication Date

9-12-2017

Document Type

Article

Disciplines

Biochemistry | Biology | Cell Biology | Cellular and Molecular Physiology | Molecular Biology

Abstract

The repeating subunit of chromatin, the nucleosome, includes two copies of each of the four core histones, and several recent studies have reported that asymmetrically-modified nucleosomes occur at regulatory elements in vivo. To probe the mechanisms by which histone modifications are read out, we designed an obligate pair of H3 heterodimers, termed H3X and H3Y, which we extensively validated genetically and biochemically. Comparing the effects of asymmetric histone tail point mutants with those of symmetric double mutants revealed that a single methylated H3K36 per nucleosome was sufficient to silence cryptic transcription in vivo. We also demonstrate the utility of this system for analysis of histone modification crosstalk, using mass spectrometry to separately identify modifications on each H3 molecule within asymmetric nucleosomes. The ability to generate asymmetric nucleosomes in vivo and in vitro provides a powerful and generalizable tool to probe the mechanisms by which H3 tails are read out by effector proteins in the cell.

Keywords

S. cerevisiae, chromatin, chromosomes, genes, synthetic biology, transcription

Rights and Permissions

Copyright © 2017, Ichikawa et al.

DOI of Published Version

10.7554/eLife.28836

Source

Elife. 2017 Sep 12;6. doi: 10.7554/eLife.28836. Link to article on publisher's site

Journal/Book/Conference Title

eLife

Comments

Full list of authors omitted for brevity. For full list see article.

Related Resources

Link to Article in PubMed

PubMed ID

28895528

Creative Commons License

Creative Commons Attribution 4.0 License
This work is licensed under a Creative Commons Attribution 4.0 License.

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